Perfume Compositions

ABSTRACT

Perfume compositions comprise between 10% and 30% in total weight of perfume ingredients selected from two groups, Group A, Group B, with the provisos that over 5% but less than 15% of the perfume composition must comprise Group A ingredients, and for compositions comprising less than 10% of Group A ingredients in the aggregate percentage of Group B ingredients present must be at least equal to the expression (2*/10−A %) where A % is the total percentage of Group A ingredients in the composition.

FIELD OF THE INVENTION

This invention relates to perfume compositions, to consumer productscontaining such perfume compositions, and to the use of such perfumecompositions to provide deodourant effects including in particularinhibiting and ameliorating the odour of urine. The invention isparticularly concerned with perfume compositions that inhibit thebacterial generation of ammonia from urea.

BACKGROUND TO THE INVENTION

The unpleasantness of urine malodour is an age-old problem. When urineis excreted into absorbent articles such as clothing, diapers orincontinence pads, or onto floors surrounding urinals or WC bowls, anammoniacal malodour may often be detected within a short time. The sameproblem occurs in pet litter, and may of course be relevant anywhere inthe house for households including cats or dogs, etc.

Urine is a clear, transparent fluid that normally has an amber color,and when fresh is generally of low odour. The average amount of urineexcreted by a human in 24 hours is about 1,200 cubic centimeters.Chemically, urine is mainly an aqueous solution of sodium chloride andorganic substances such as urea and uric acid. Normally, it containsabout 960 parts of water to 40 parts of solid matter. Many hundreds ofdifferent mineral salts and organic compounds are present in urine,albeit at trace levels for a significant proportion of these. The pH ofnormal urine is between 4.5 and 7.8, but usually it ranges between 5.0and 6.0, due to obligatory excretion of acid produced every day.

The major components of urine malodour are: ammonia, volatile fattyacids (primarily acetic, propionic, butyric, formic); volatile sulphurcompounds e.g hydrogen sulfide (H₂S) and methyl sulphides such as methylmercaptan (CH₃SH); other nitrogenous compounds such as indole, skatole,pyridine, pyrrole, ethylamine; various other volatiles including benzylalcohol, phenol, p-cresol, ethanol, methanol, acetone, methyl ethylketone, acetaldehyde, propionaldehyde, pentanone, heptanone, propanol,butanol, octanol. These odourous molecules are mainly produced as aresult of the bacterial degradation of exogenous materials such as ureaand uric acid found in urine, though trace levels of certain materialsmay reflect materials found in the diet or in the environment. Thenitrogenous bases, in particular ammonia, contribute significantly tothe malodour recognised by most people from used diapers or otherhygiene products such as adult incontinence products. This malodourarises at least partly from the bacterially mediated degradation ofurea, from the metabolism of microorganisms present on the skin or fromthe urogenital tract, for example from the growth of Proteus andMicrococcus species. All strains of Proteus spp. form the enzyme ureaseduring their metabolism. Urease has the ability to rapidly break downurea (constituting about 2% of human urine) into ammonia causingunpleasant odour. The headspace composition above stale urine comprisesa variety of materials, but the dominant malodour contributor under mostconditions is invariably ammonia. Techniques that lead to reduction inthe amount of ammonia present above urine are therefore of possibleutility in product sectors associated with sanitation, hygiene, andincontinence. Several approaches are known in the art that address thisneed.

Antimicrobial agents used in personal products are designed to reducethe population, inhibit the growth or diminish the metabolic activitiesof microorganisms associated closely with the body—on the surface of theskin, in mucosal surfaces, in the urogenital tract, etc. Typical agentsof this nature include triclosan (2′,4,4′-trichloro-2-hydroxydiphenylether) and zinc oxide which are well known to exert antimicrobial anddeodourant effects. The use of common deodourant actives results in anon-selective antimicrobial action exerted upon most of the skin'snatural microflora. This can represent an undesirable feature of suchdeodourant formulations, since the natural microflora provides aprotective barrier (colonisation resistance) against invasion bypotentially pathogenic bacteria. Certain perfume components and mixturesthereof may contribute to such antimicrobial effects. For example,published US application US2004266302 relates to a disposable absorbentarticle containing an encapsulated antimicrobial essential oil for odourcontrol.

PCT publication WO 2002/47472 relates to products and methods thatutilise a urease inhibitor formed from a polyanionic, and preferablyamine-based, chelating agent and a divalent heavy metal ion, to preventor minimize ammonia odour produced by the degradation of urea insecreted or excreted body fluids. Similarly WO 97/46187 relates toabsorbent articles in particular sanitary napkins and panty linershaving an odour control system comprising a polyfunctionally substitutedaromatic chelating agent for improved odour control.

Much of the art is concerned with the use of odour absorbent materials.For example, WO 2001/80915 relates to absorbent articles that comprise acationic polysaccharide, preferably chitosan material, and silicate.These articles claim to deliver improved odour control performance(synergistic odour reduction) and improved fluid handlingproperties/absorption performance. WO 94/25077 relates to odour controlthrough an absorbent article containing a boric acid/sodium tetraboratebuffer. EP 509409 relates to malodour control though the design of anabsorbent article containing a deodourizing blend of anhydrous,non-buffer blend of at least basic and pH neutral odour absorbingparticles. U.S. Pat. No. 6,031,147 discloses an absorbent productcomprising a hydrogel-forming polymeric absorbent material and asurface-active agent such as ethoxylated sorbitan monooleate, having ahydrophilic/lipophilic balance of less than about 12.

WO 99/06078 describes absorbent materials containing cyclodextrin as anodour control material. WO 98/26808 describes odour control provided bya combination of a material that inhibits the formation of odour (andhas at least one attribute selected from the group consisting ofantimicrobial activity, urease inhibition activity, pH adjustmentactivity) and an odour-absorbing material for objectionable odourmolecules selected from the group consisting of cyclodextrin, zeolite,activated carbon, kieselguhr, acid salt forming materials and mixturesthereof. The scent signal is provided by cyclodextrin/perfume inclusioncomplexes and/or matrix perfume microcapsules to assure the wearer thatthe product is working.

WO 2000/51652 describes the use of oxidising agents such as a peroxyacidin combination with an odour-absorbing agent such as silica and/orzeolite. WO2003/051413 and WO 2003/051410 relate to a fibrous absorbentmaterial or cellulose fibers treated with a carboxylic acid based odourcontrol agent.

Perfumes have long been recognised as beneficial in hygiene and sanitarysectors. WO 98/25562 describes a diaper design that contains perfumezones and microcapsules as release agents to provide odour control.Published application US 2003/072733 describes a process for absorbingmoisture and/or malodour while providing a fragrance to the surroundingambience. WO 2005/044320 relates to a dual purpose volatile substancecontrolling composition comprising a sorbent and a fragrance componentdesigned to control malodours including those resulting from bodilyfluids. The design involves volatile substance sorption directly linkedto fragrance release.

Perfumes may simply mask malodours. WO 2004/10325 describes sanitaryabsorbent articles comprising a non-aqueous volatile cooling agent suchas menthyl lactate or perfume. WO 2004/108177 describes theincorporation of a starch-encapsulated accord into products thatreleases perfume to minimise odour. However, perfume compositions havebeen disclosed which exhibit effective deodourant action for specificmalodours. For example, WO 2000/01356 describes certain perfumecomponents and compositions thereof, useful in reducing or preventingbody malodour. The perfume components (or compositions comprising theperfume components) are described as inhibiting coryneform bacteria thatare capable of catabolising fatty acids and are responsible for theproduction of short chain fatty acid malodour. In this way, the perfumecomponents (or compositions thereof) in-use produce a deodourant effect.However, many of the deodourant perfumes disclosed in the art haverelatively high odour intensities that are unsuitable for use in thesanitary or incontinence product sector, and/or are not effective incounteracting or inhibiting ammonia malodour.

In spite of the above mentioned disclosures there still exists a needfor cost-effective products that combat urine-derived malodour moreefficiently, both from the perspective of malodour prevention as well asmalodour amelioration, and do not suffer from the potentialdisadvantages of exploiting highly antimicrobial actives, whilstbenefiting from the presence of perfumes that enjoy broad consumeracceptability.

It has now been surprisingly found that particular perfume compositionsare capable of inhibiting the development of ammonia from urea atsub-minimum inhibitory growth concentrations (MIC), and are effective incounteracting urine malodour in spite of exhibiting relatively lowperfume odour intensities.

SUMMARY OF THE INVENTION

In one aspect, this invention provides a perfume composition thatinhibits the formation of urine malodour, and is effective inameliorating the perception of urine malodour if present. Such a perfumecomposition comprises between 10% and 30% in total by weight of perfumeingredients selected from two groups:

Group A, consisting of (3Z)-hex-3-enyl acetate;3-(1,3-benzodioxol-5-yl)-2-methylpropanal; (3Z)-hex-3-en-1-ol;1-[4-(methyloxy)phenyl]ethanone; 3-methyl-5-phenylpentan-1-ol;5-heptyldihydrofuran-2(3H)-one; phenylmethyl acetate;2-(4-methylcyclohex-3-en-1-yl)propan-2-ol;(3E)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one;(3E)-4-(2,6,6-trimethylcyclohex-1-en-1-yl)but-3-en-2-one;4-hydroxy-3-(methyloxy)benzaldehyde;1-methyl-3-(2-methylpropyl)cyclohexanol;7,9-dimethylspiro[5.5]undecan-3-one; perfume 1 being Lavandin AB8381™;methyl 2,4-dihydroxy-3,6-dimethylbenzoate; 3,7-dimethyloctan-1-ol;2-(methyloxy)-4-propylphenol; perfume 2 being Rosenta AB8428™;1-{[(1R,2S)-2-(1,1-dimethylethyl)cyclohexyl]oxy}butan-2-ol; perfume 3being Headspace Freesia AB7254A™; 5-hexyldihydrofuran-2(3H)-one;prop-2-enyl[(2-methylbutyl)oxy]acetate; 1,3-benzodioxole-5-carbaldehyde;[4-(1-methylethyl)cyclohexyl]methanol; 2-hexylcyclopent-2-en-1-one;methyl (2E)-3-phenylprop-2-enoate; 2,6-dimethyloct-7-en-2-ol;2-methyl-3-[4-(1-methylethyl)phenyl]propanal; andGroup B consisting of 2-phenylethanol; 3,7-dimethyloctan-3-ol(tetrahydro linalol), 2-(methyloxy)-4-[(1E)-prop-1-enyl]phenyl acetate;4-(methyloxy); (2E)-1-(2,6,6-trimethylcyclohex-2-en-1-yl)but-2-en-1-one;perfume 4 being Bergamot AB8392™,(3E)-3-methyl-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one;cyclopentadecanone; cyclohexadecanolide; prop-2-enyl3-cyclohexylpropanoate; 3-[3-(1-methylethyl)phenyl]butanal;(3Z)-hex-3-enyl methyl carbonate;(1-methyl-2-{[1S,3R,5R)-1,2,2-trimethylbicyclo[3.1.0]hex-3-yl]methyl}cyclopropyl)methanol;[3,3-bis(methyloxy)propyl]benzene; perfume 5 being Coumarex I Mod™;tricyclo[5.2.1.0^({2,6})]dec-4-en-8-yl propanoate;with the provisos that over 5% but less than 15% of the perfumecomposition must comprise Group A ingredients, and for compositionscomprising less than 10% of Group A ingredients the percentage of GroupB ingredients present must be at least equal to the expression 2*(10−A%) where A % is the total percentage of Group A ingredients in thecomposition.

In another aspect, the invention relates to a method of preventing orameliorating urine malodour comprising bringing into contact with urineor urine residues an effective amount of a perfume composition accordingto the invention.

At least 0.3 wt % of a perfume ingredient must be present before it maybe considered to contribute significantly towards the efficacy of thecompositions, i.e. perfume ingredients present at concentrations below0.3 wt % and ignored in the calculation of the number of Group A andGroup B ingredients in the composition.

Preferred perfume compositions comprise at least three Group Aingredients, more preferably at least four ingredients and mostpreferably at least six ingredients.

For the purposes of this invention a perfume composition is defined as amixture of perfume ingredients, if desired mixed with or dissolved in asuitable solvent or solvents and/or mixed with a solid substrate.Perfume ingredients are well known to those skilled in the art, andinclude those mentioned, for example, in S. Arctander, Perfume andFlavor Chemicals (Montclair, N.J., 1969), in S. Arctander, Perfume andFlavor Materials of Natural Origin (Elizabeth, N.J., 1960) and in“Flavor and Fragrance Materials—1991”, Allured Publishing Co. Wheaton,Ill. USA. Perfume ingredients may include natural products such asextracts, essential oils, absolutes, resinoids, resins, concretes etc.,and also synthetic substances such as hydrocarbons, alcohols, aldehydes,ketones, ethers, acids, esters, acetals, ketals, nitriles, etc.,including saturated and unsaturated compounds, aliphatic, macrocyclicand heterocyclic compounds.

References herein to the percentage by weight of perfume ingredientsmeans relative to the total weight of perfume ingredients in the perfumecomposition and includes materials that are used within perfumery asvehicles or solvents for other perfume ingredients, for exampledipropylene glycol, isopropyl myristate, benzyl benzoate, diethylphthalate, triacetin and triethyl citrate.

Perfumes constructed according to the above design provide effectiveurease inhibition without being overtly strongly antimicrobial (asindicated by tests described below) and also are able to counteracturine malodour olfactorily.

In a further aspect, the invention provides a perfumed consumer productcomprising a perfume composition in accordance with the invention. Forthe purposes of this invention a consumer product means comprises asolid, liquid or soft solid formulation especially for use in or on asubstrate such as skin, hair (including fur), clothing or hard surface.Examples of such consumer products include bathroom and kitchencleaners, carpet cleaners, polishes, personal body refreshers anddeodourants, pet deodourants, in a variety of formats such as liquids(particularly as delivered by trigger sprays or aerosols), gels andpowders, all of which are well known in the art. Another example of aconsumer product relevant to the present invention is pet litter.

In such consumer products as little as 0.1% by weight of the perfumecomposition in the product will suffice.

The invention also covers use of a perfume composition according to theinvention for the purpose of inhibiting urea breakdown to form ammonia.Preferred features of this aspect are as discussed below in connectionwith the perfume composition of the invention. A concomitant effect ofthe inhibition of ammonia production is that pH remains relativelyconstant, or at least the rate of increase of pH is much lower than inthe absence of the perfume composition. Such pH control may be ofindirect benefit in areas other than malodour management.

In a further aspect, the invention provides an article suitable forpreventing or ameliorating urine malodour, comprising an effectiveamount of a perfume composition according to the invention. Sucharticles of manufacture include diapers, incontinence pads, hygienicbody wipes, and catamenials including sanitary pads and sanitary towels.Perfume compositions of the invention may be incorporated into or ontosuch articles by any suitable means known in the art, for example bybringing them into contact with adsorbents present in such articles,although they be used in association with a wide variety of elements ofsuch articles. It may sometimes be advantageous to encapsulate theperfumes of the invention prior to incorporation into such articles.

Perfume Ingredients of the Invention

Perfumes formulated to the guidelines described herein will maintain thepH of a urea-supplemented microbial suspension (as described below inExample 1) at a pH that is at least 1.2 pH units lower than that of anon-perfumed control. It is believed that this difference in pH isdirectly correlated to a decrease in ammonia production. It is highlydesirable that the effect is achieved at sub-inhibitory growth levels.Two groups of perfume ingredients have been identified. Group Amaterials have the highest efficacy, but Group B may be used in place ofa fraction of the Group A materials where this is desirable in order toachieve the right balance of hedonic properties, anti-microbial actionand sensory-derived malodour counteraction.

Group A ingredients are listed below, where names in parenthesesrepresents equivalent names—either trivial names commonly used withinthe fragrance and flavour industry, or tradenames that are sources forthe material cited.

Group A Materials:

-   (3Z)-hex-3-enyl acetate (cis-3-hexenyl acetate),-   3-(1,3-benzodioxol-5-yl)-2-methylpropanal[Helional™ (IFF)],-   (3Z)-hex-3-en-1-ol (cis-3-hexenol)-   1-[4-(methyloxy)phenyl]ethanone (paramethoxy acetophenone),-   3-methyl-5-phenylpentan-1-ol[Mefrosol™ (G)],-   5-heptyldihydrofuran-2(3H)-one (undecalactone gamma),-   phenylmethyl acetate (benzyl acetate),-   2-(4-methylcyclohex-3-en-1-yl)propan-2-ol (terpineol alpha),-   (3E)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one (ionone    alpha),-   (3E)-4-(2,6,6-trimethylcyclohex-1-en-1-yl)but-3-en-2-one (ionone    beta),-   4-hydroxy-3-(methyloxy)benzaldehyde (vanillin),-   1-methyl-3-(2-methylpropyl)cyclohexanol[Rossitol™ (G)],-   7,9-dimethylspiro[5.5]undecan-3-one[Dispirone™ (G)],-   perfume 1 being Lavandin AB8381™,-   methyl 2,4-dihydroxy-3,6-dimethylbenzoate (moss oakmoss synthetic),-   3,7-dimethyloctan-1-ol (tetrahydrogeraniol),-   2-(methyloxy)-4-propylphenol (dihydroeugenol),-   perfume 2 being Rosenta AB842™,-   1-{[(1R,2S)-2-(1,1-dimethylethyl)cyclohexyl]oxy}butan-2-ol[Amber    Coretm (G)],-   perfume 3 being Headspace Freesia AB7254A™,-   5-hexyldihydrofuran-2(3H)-one (decalactone gamma),-   prop-2-enyl[(2-methylbutyl)oxy]acetate (Allyl amyl glycolate),-   1,3-benzodioxole-5-carbaldehyde (heliotropin),-   [4-(1-methylethyl)cyclohexyl]methanol[Mayol™ (F)],-   2-hexylcyclopent-2-en-1-one (iso-jasmone),-   methyl (2E)-3-phenylprop-2-enoate (methyl cinnamate),-   2,6-dimethyloct-7-en-2-ol (dihydromyrcenol),-   2-methyl-3-[4-(1-methylethyl)phenyl]propanal (Cyclamen aldehyde).    Group B ingredients:-   2-phenylethanol (phenyl ethyl alcohol);-   3,7-dimethyloctan-3-ol (tetrahydrolinalol);-   2-(methyloxy)-4-[(1E)-prop-1-enyl]phenyl acetate (isoeugenyl    acetate);-   4-(methyloxy)benzaldehyde (anisic aldehyde);-   (2E)-1-(2,6,6-trimethylcyclohex-2-en-1-yl)but-2-en-1-one (damascone    alpha);-   perfume 4 being Bergamot AB8392™;-   (3E)-3-methyl-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one    (Methyl ionone alpha iso);-   Cyclopentadecanone[Silvanone™ (G)];-   Oxacycloheptadecan-2-one (Silvanone™ (G), cyclohexadecanolide);-   prop-2-enyl 3-cyclohexylpropanoate (allyl cyclo hexyl propionate);-   3-[3-(1-methylethyl)phenyl]butanal[Florohydral™ (G)];-   (3Z)-hex-3-enyl methyl carbonate (cis-3-hexenyl methyl carbonate);-   (1-methyl-2-{[(1S,3R,5R)-1,2,2-trimethylbicyclo[3.1.0]hex-3-yl]methyl}cyclopropyl)methanol[Javanol™    (G)];-   [3,3-bis(methyloxy)propyl]benzene (dimethyl hydro cinnamyl);-   perfume 5 being Conmarex I Mod™ (IFF);-   tricyclo[5.2.1.0^({2,6})]dec-4-en-8-yl propanoate[Florocyclene™    (G)].

Key: G=Givaudan; IFF=International Flavours and Fragrances; F=Firmenich

wherein perfumes 1 to 5 have the following compositions:

Per- Per- Per- Per- Per- Ingredient fume 1 fume 2 fume 3 fume 4 fume 5Allylamyl glycolate 0.8 Anisaldehyde 0.3 Benzophenone 2.0 Borneol 1.0Camphene 0.8 Camphor 9.2 Caryophyllene 4.5 Cedarwood Texan oil 0.8Cineole 7.0 Citral Diethylacetal 1.5 Citronellyl formate 1.8 Citronellylpropionate 2.5 Dihydrolinalol 6.5 Dihydro myrcenyl 11.0 acetate DihydroTerpinyl 13.5 31.5 Acetate 2,6-Dimethylheptan-2-ol 2.0 Dipropyleneglycol 5.0 88.0 Geranylacetate 5.0 Geranylacetone 1.0 Geranyl formate1.2 Hercolyn DE (TM) 4.5 gamma-hexalone 5.6 beta-ionone 3.0 24.5 Linalylacetate 1.0 32.0 Mefrosol (TM) 10.0 14.0 3-Methylbut-2-enyl 4.5 benzoateNeryl acetate 1.0 Ocimene 1.0 para-tert-butyl- 4.5 cyclohexyl acetatePhenyl acetaldehyde 2.0 dimethylacetal 2-phenylethyl alcohol 27.02-phenylethyl 24.0 phenylacetate 2-phenylethyl 1.0 salicylatealpha-pinene 1.4 beta-pinene 6.8 Clary sage oil 0.8 gamma-terpinene 6.0alpha-terpineol 4.2 12.2 Terpinyl acetate 17.9 alpha-terpinyl 2.5isobutyrate Tetrahydrogeraniol 1.4 Tetrahydrolinalol 60.3 4.0Tetrahydrolinalyl 4.5 acetate Minor components 6.8 5.3 3.0 4.8 1.6Totals 100 100 100 100 100

KEY

Mefrosol is 3-methyl-5-phenylpentan-1-olHercolyn DE is a mixture of methyl dihydroabietate andtetrahydroabietate

Particularly preferred Group A materials are:

-   (3Z)-hex-3-enyl acetate; 3-(1,3-benzodioxol-5-yl)-2-methylpropanal;    (3Z)-hex-3-en-1-ol; 3-methyl-5-phenylpentan-1-ol;    5-heptyldihydrofuran-2(3H)-one; phenylmethyl acetate;    2-(4-methylcyclohex-3-en-1-yl)propan-2-ol;    2,6-dimethyloct-7-en-2-ol;    (3E)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one;    (3E)-4-(2,6,6-trimethylcyclohex-1-en-1-yl)but-3-en-2-one;    4-hydroxy-3-(methyloxy)benz aldehyde;    1,3-benzodioxole-5-carbaldehyde.

It has also been discovered that the presence of different perfumesolvents within the composition may affect activity slightly.Embodiments incorporating triethyl citrate are preferred.

EXAMPLES Example 1 MIC Estimation for Fragrance and FragranceIngredients

The minimum inhibitory concentration of perfumes may be determined bythe following method.

Stock solutions (8% w/w) of perfume were prepared in sterile tryptonesoya broth.

Test Strain:

Proteus vulgaris NCTC 4175 (National Collection of Type Cultures, PublicHealth Laboratory Service, Central Public Health Laboratory, 61Colindale Avenue, London)

Cultures were grown in 10 ml of tryptone soya broth (TSB) (Oxoid,Basingstoke, UK), for 16-24 hours, in shaken flasks at 37° C. Thecultures were then diluted in sterile 0.1% special peptone solution(Oxoid, Basingstoke, UK) to give a concentration of bacteria ofapproximately 10⁶ colony-forming units (cfu) per ml.

Test ingredients were diluted in sterile TSB. For each test bacterialculture, each row of a standard, 96-well plastic microtitre plate(labelled A-H) was allocated to one sample, thus eight samples perplate. Row H contained only TSB for use as a bacterial control toindicate the degree of turbidity resulting from bacterial growth in theabsence of any test material. Aseptically, 200 μl of the initialdilution of ingredient was transferred to the 1st and 7th well of theappropriate row. All other test wells were filled with 100 μl of sterileTSB using an 8-channel micropipette. The contents of each of the wellsin column 1 were mixed by sucking samples up and down in pipette tips,before 100 μl was transferred to column 2 The same sterile pipette tipswere used to transfer 100 μl of each well in column 7, into theappropriate well in column 8. This set of eight tips was then discardedinto disinfectant solution. Using eight fresh, sterile tips the processwas repeated by transferring 100 μl from column 2 into column 3 (and 8into 9). The process was continued until all wells in columns 6 and 12contained 200 μl. After mixing, 100 μl was discarded from wells incolumns 6 and 12 to waste. Finally, 100 μl of pre-diluted bacterialculture (approx. 106 cfu/ml) was added, thus giving 200 μl final volumein each well.

A blank plate was prepared for each set of eight samples in exactly thesame way, except that 100 μl of sterile 0.1% special peptone was addedinstead of bacterial culture. Test and control plates were sealed usingautoclave tape and incubated for 18 hours at 37° C.

The microtitre plate reader (Thermo Multiskan Ascent) was preset togently agitate the plates, to mix the contents. The absorbance at 540 nmwas used as a measure of turbidity resulting from bacterial growth. Bothcontrol (un-inoculated plate) and inoculated plates were read for eachset of samples. The absorbance readings from the control plate weresubtracted from the relevant inoculated plate readings thus removingturbidity due to perfume and possible colour changes during incubation.Thus the corrected readings generated were absorbances resulting fromturbidity from bacterial growth. The MIC was taken as the concentrationof ingredient required to inhibit growth so that the average change inabsorbance during the incubation period was <0.3.

Example 2 Urease Assay—Estimation of Fragrance and Fragrance IngredientsEffect on Ammonia Production

The method adopted was based on the rapid method devised by C. A.Stuart, Elizabeth van Stratum and Robert Rustigan Further Studies onUrease Production by Proteus and Related Organisms J. Bacteriol. 1945,49: 437-444.

SSR Medium: 380 ml distilled water, 364 mg KH₂PO₄, 380 mg Na₂HPO₄, 8 gUrea, 40 mg Yeast extract, 20 ml 0.02% phenol red indicator, pH 6.8,solution filter sterilised. Culture: Liquid culture of P. vulgarisNCTC4175 was prepared by adding loops of fresh culture from solid media(Tryptone Soya Agar) to sterile 0.1% special peptone solution.Absorbance of culture was adjusted to be OD610 nm>2.3

Fragrances were aseptically prepared at 2500 ppm in medium in duplicate(5 ml volumes) using solubiliser Synperonic 91/10 at ratio of 1:2fragrance:solubiliser. In addition 5 ml volumes of positive and negativecontrols were prepared. Positive controls and all test solutions wereinoculated by adding 200 μl of prepared culture, mixed and incubated for24 hours @ 37oC. No culture was added to the negative growth controls.Initially solutions were orange in colour. Urease activity can be notedby the production of a purple/red colour, indicating high pH resultingfrom ammonia production. The degree of urease activity/ammoniaproduction was assessed by pH electrode.

Example 3 Perfume Compositions

Fragrance formulations 3.2, 3.4, and 3.8 detailed in Table 1 werecreated following the creative guidelines of the present invention.Formulations, 3.1, 3.3, 3.5, 3.6 and 3.7 are comparatives that falloutside these guidelines. All examples bar timbers 3.5 and 3.6 meet theUrease Inhibition Assay target of a difference of 1.2 pH units comparedto the positive control. However only Formulations 3.2, 3.4 and 3.8achieve this effect at sub-MIC level. These three perfumes were alsoevaluated in a urine malodour test as described in example 3 ofPCT/GB2007/001172 using malodour model A of example 1 therein. All threewere found to be effective in reducing the perception of the urinemalodour.

TABLE 1 Perfume Formulations (% w/w) Ingredient 3.1 3.2 3.3 3.4 3.5 3.63.7 3.8 Acetyl tributyl citrate 10 10 5 5 Allyl amyl glycolate (A) 1 3Amyl salicylate 5 5 4 7 Anisic aldehyde (B) 5 4 0.5 Bangalol^(tm) (G)0.3 0.3 Benzyl acetate (A) 1 cis-3-hexenol (A) 0.5 0.5 cis-3-hexenylsalicylate 1 coumarex I mod^(tm) (B) 5 Cyclamen aldehyde (A) 5Decalactone gamma (A) 0.5 Dihydromyrcenol (A) 10 5 Dispirone^(tm) (G)(A) 0.1 0.5 Ethylene brassylate 10 15 10 14.5 12 12 Ethyl safranate^(tm)(G) 0.2 Florosa^(tm) (G) 0.2 0.5 Habanolide^(tm) (F) 1 2 3 Helional^(tm)(A) 1 1 1 1 2 Heliotropin (A) 3 1 1 2 Hexyl salicylate 5 12 10 12 10 15Ionone alpha (A) 2 Mayol^(tm) (F) (A) 1 Mefrosol^(tm) (G) (A) 0.9 1 1 12.5 Methyl dihydrojasmonate 8 14 15 15 10 8 para tert butyl cyclo hexyl5 acetate Moss oakmoss synthetic (A) 0.1 Phenoxyethanol 5 10 5 Phenylethyl alcohol (B) 10 7.5 5 6.5 7 10 Rossitol^(tm) (G) (A) 2Silvanone^(tm) (G) (B) 1 Terpineol alpha (A) 6 5 2 Tetrahydrogeraniol(A) 1.5 3 Tetrahydrolinalol (B) 5 2 1 1 3 10.5 Triethyl citrate 34 36 5033 48 50 45 50 Undecalactone gamma (A) 0.5 0.5 Vanillin (A) 2 1.9 2 1Total Group A ingredients 20 5.5 15 7 5 0 14.5 10 Total Group Bingredients 16 19.5 10 8 10 0 20.5 0 Total Group A + Group B 36 25 25 1515 0 35 10 KEY: (A) = group A material: (B) = Group B material (G) =Givaudan; (F) = Firmenich

Following the methods outlined previously, the above examples giveresults detailed in Table 2.

TABLE 2 Results of MIC and Urease inhibition assays Urease InhibitionAssay MIC assay Results at 2500 ppm MIC vs pH unit difference Pr.vulgaris Fragrance Average pH from +ve control (ppm) Example 3.1 7.501.60 2500 Example 3.2 7.85 1.25 5000 Example 3.3 7.69 1.41 1250 Example3.4 7.67 1.43 10000 Example 3.5 8.44 0.66 10000 Example 3.6 8.78 0.3210000 Example 3.7 7.64 1.46 2500 Example 3.8 7.58 1.52 5000 +ve control9.10 N/A N/A −ve control 6.99 N/A N/A

1. (canceled)
 2. (canceled)
 3. (canceled)
 4. (canceled)
 5. (canceled) 6.(canceled)
 7. (canceled)
 8. (canceled)
 9. (canceled)
 10. A method ofinhibiting the formation of ammonia from urea, the method comprising thestep of: contacting urine or urine residues with a perfume compositioncomprising between 10% and 30% in total by weight of perfume ingredientsselected from two groups: Group A, consisting of: (3Z)-hex-3-enylacetate; 3-(1,3-benzodioxol-5-yl)-2-methylpropanal; (3Z)-hex-3-en-1-ol;1-[4-(methyloxy)phenyl]ethanone; 3-methyl-5-phenylpentan-1-ol;5-heptyldihydrofuran-2(3H)-one; phenylmethyl acetate;2-(4-methylcyclohex-3-en-1-yl)propan-2-ol;(3E)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one;(3E)-4-(2,6,6-trimethylcyclohex-1-en-1-yl)but-3-en-2-one;4-hydroxy-3-(methyloxy)benzaldehyde;1-methyl-3-(2-methylpropyl)cyclohexanol;7,9-dimethylspiro[5.5]undecan-3-one; perfume 1 being LAVANDIN AB8381;methyl 2,4-dihydroxy-3,6-dimethylbenzoate; 3,7-dimethyloctan-1-o1;2-(methyloxy)-4-propylphenol; perfume 2 being ROSENTA AB8428;1-{[(1R,2S)-2-(1,1-dimethylethyl)cyclohexyl]oxy}butan-2-ol; perfume 3being HEADSPACE FREESIA AB7254A; 5-hexyldihydrofuran-2(3H)-one;prop-2-enyl[(2-methylbutyl)oxy]acetate; 1,3-benzodioxole-5-carbaldehyde;[4-(1-methylethyl)cyclohexyl]methanol; 2-hexylcyclopent-2-en-1-one;methyl (2E)-3-phenylprop-2-enoate; 2,6-dimethyloct-7-en-2-ol;2-methyl-3-[4-(1-methylethyl)phenyl]propanal; and, Group B consistingof: 2-phenylethanol; 3,7-dimethyloctan-3-ol (tetrahydro linalol),2-(methyloxy)-4-[(1E)-prop-1-enyl]phenyl acetate;4-(methyloxy)benzaldehyde;(2E)-1-(2,6,6-trimethylcyclohex-2-en-1-yl)but-2-en-1-one; perfume 4being BERGAMOT AB8392,(3E)-3-methyl-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one;cyclopentadecanone; cyclohexadecanolide; prop-2-enyl3-cyclohexylpropanoate; 3-[3-(1-methylethyl)phenyl]butanal;(3Z)-hex-3-enyl methyl carbonate;(1-methyl-2-{[(1S,3R,5R)-1,2,2-trimethylbicyclo[3.1.0]hex-3-yl]methyl}cyclopropyl)methanol;[3,3-bis(methyloxy)propyl]benzene; perfume 5 being COUMAREX I Mod;tricyclo[5.2.1.0.sup. {2,6}]dec-4-en-8-yl propanoate; with the provisosthat over 5% but less than 15% of the perfume composition must compriseGroup A ingredients, and for compositions comprising less than 10% ofGroup A ingredients, the aggregate percentage of Group B ingredientspresent must be at least equal to the expression 2*(10−A %) where A % isthe total percentage of Group A ingredients in the composition.
 11. Themethod according to claim 10, wherein the perfume composition comprisesat least 3 perfume ingredients of Group A.
 12. The method according toclaim 11, wherein the perfume composition comprises at least 4 perfumeingredients of Group A.
 13. The method according to claim 12, whereinthe perfume composition comprises at least 6 perfume ingredients ofGroup A.
 14. The method according to claim 10, wherein the perfumeingredients of Group A are selected from the group consisting of:(3)-hex-3-enyl acetate; 3-(1,3-benzodioxol-5-yl)-2-methylpropanal;(3Z)-hex-3-en-1-ol; 3-methyl-5-phenylpentan-1-ol;5-heptyldihydrofuran-2(3H)-one; phenylmethyl acetate;2-(4-methylcyclohex-3en-1-yl)propan-2-ol; 2,6-dimethyloct-7en-2-ol;(3E)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one;(3E)-4-(2,6,6-trimethylcyclohex-1-en-1-yl)but-3 en-one;4-hydroxy-3-(methyloxy)benzaldehyde; 1,3-benzodioxole-5-carbaldehyde.15. A method according to claim 10, wherein the said perfume compositionis incorporated in an article adapted for preventing or amelioratingurine malodour.
 16. A method according to claim 15, wherein the saidarticle is a catamenial.
 17. A method according to claim
 15. wherein thesaid article is adapted to control incontinence.